Urea is one of the dominant organic nitrogenous compounds in the oligotrophic oceans. Compared to the knowledge of nitrogen transformation of nitrogen fixation, ammonia oxidization, nitrate and nitrite reduction mediated by sponge-associated microbes, our knowledge of urea utilization in sponges and the phylogenetic diversity of sponge-associated microbes with urea utilization potential is very limited. In this study, Marinobacter litoralis isolated from the marine sponge Xestospongia testudinaria and the slurry of X. testudinaria were found to have urease activity. Subsequently, phylogenetically diverse bacterial ureC genes were detected in the total genomic DNA and RNA of sponge X. testudinaria, i.e., 19 operative taxonomic units (OTUs) in genomic DNA library and 8 OTUs in cDNA library at 90% stringency. Particularly, 6 OTUs were common to both the genomic DNA library and the cDNA library, which suggested that some ureC genes were expressed in this sponge. BLAST and phylogenetic analysis showed that most of the ureC sequences were similar with the urease alpha subunit of members from Proteobacteria, which were the predominant component in sponge X. testudinaria, and the remaining ureC sequences were related to those from Magnetococcus, Cyanobacteria, and Actinobacteria. This study is the first assessment of the role of sponge bacterial symbionts in the regenerated utilization of urea by the detection of transcriptional activity of ureC gene, as well as the phylogenetic diversity of ureC gene of sponge bacterial symbionts. The results suggested the urea utilization by bacterial symbionts in marine sponge X. testudinaria, extending our understanding of nitrogen cycling mediated by sponge-associated microbiota.

Actinobacteria are widely distributed in the marine environment. To date, few studies have been performed to explore the coral-associated Actinobacteria, and little is known about the diversity of coral-associated Actinobacteria. In this study, the actinobacterial diversity associated with one soft coral Alcyonium gracllimum and one stony coral Tubastraea coccinea collected from the East China Sea was investigated using both culture-independent and culture-dependent approaches. A total of 19 actinobacterial genera were detected in these two corals, among which nine genera (Corynebacterium, Dietzia, Gordonia, Kocuria, Microbacterium, Micrococcus, Mycobacterium, Streptomyces, and Candidatus Microthrix) were common, three genera (Cellulomonas, Dermatophilus, and Janibacter) were unique to the soft coral, and seven genera (Brevibacterium, Dermacoccus, Leucobacter, Micromonospora, Nocardioides, Rhodococcus, and Serinicoccus) were unique to the stony coral. This finding suggested that highly diverse Actinobacteria were associated with different types of corals. In particular, five actinobacterial genera (Cellulomonas, Dermacoccus, Gordonia, Serinicoccus, and Candidatus Microthrix) were recovered from corals for the first time, extending the known diversity of coral-associated Actinobacteria. This study shows that soft and stony corals host diverse Actinobacteria and can serve as a new source of marine actinomycetes.

Compared with the knowledge of sponge-associated bacterial diversity and ecological roles, the fungal diversity and ecological roles of sponges remain largely unknown. In this study, the fungal diversity and protein synthesis potential in two South China Sea sponges Theonella swinhoei and Xestospongia testudinaria were investigated by rRNA vs. rRNA gene analysis. EF4/fung5 was chosen after a series of PCR tests to target fungal 18S rRNA and 18S rRNA gene. Altogether, 283 high-quality sequences were obtained, which resulted in 26 Operational taxonomic units (OTUs) that were assigned to Ascomycota, Basidiomycota, and Blastocladiomycota. At subphylum level, 77.3% of sponge-derived sequences were affiliated with Pezizomycotina. The fungal compositions of T. swinhoei and X. testudinaria were different from that of ambient seawater. The predominant OTU shared between two sponges was rare in seawater, whereas the most abundant OTUs in seawater were not found in sponges. Additionally, the major OTUs of sponge cDNA datasets were shared in two sponges. The fungal diversity illustrated by sponge cDNA datasets correlated well with that derived from sponge DNA datasets, indicating that the major members of sponge-associated fungi had protein synthesis potential. This study highlighted the diversity of Pezizomycotina in marine sponge-fungi symbioses and the necessity of investigating ecological roles of sponge-associated fungi.